EP1212323A2 - Pyridino pentaazamacrocyle substitue possedant une activite de superoxyde dismutase - Google Patents

Pyridino pentaazamacrocyle substitue possedant une activite de superoxyde dismutase

Info

Publication number
EP1212323A2
EP1212323A2 EP00966722A EP00966722A EP1212323A2 EP 1212323 A2 EP1212323 A2 EP 1212323A2 EP 00966722 A EP00966722 A EP 00966722A EP 00966722 A EP00966722 A EP 00966722A EP 1212323 A2 EP1212323 A2 EP 1212323A2
Authority
EP
European Patent Office
Prior art keywords
ofthe
carbon atoms
mmol
substituted
ligand
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP00966722A
Other languages
German (de)
English (en)
Other versions
EP1212323B1 (fr
Inventor
Dennis Riley
William L. Neumann
Susan L. Henke
Patrick Lennon
Karl W. Aston
Daniela Salvemini
James A. Sikorski
Yvette Fobian
Margaret L. Grapperhaus
Carrie L. Kusturin
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Monsanto Co
Original Assignee
Monsanto Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Monsanto Co filed Critical Monsanto Co
Priority to EP04003746A priority Critical patent/EP1420022B1/fr
Priority to EP04003751A priority patent/EP1420019B1/fr
Publication of EP1212323A2 publication Critical patent/EP1212323A2/fr
Application granted granted Critical
Publication of EP1212323B1 publication Critical patent/EP1212323B1/fr
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/06—Peri-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/555—Heterocyclic compounds containing heavy metals, e.g. hemin, hematin, melarsoprol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
    • A61P25/36—Opioid-abuse
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/02—Non-specific cardiovascular stimulants, e.g. drugs for syncope, antihypotensives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/08—Bridged systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F13/00—Compounds containing elements of Groups 7 or 17 of the Periodic Table
    • C07F13/005—Compounds without a metal-carbon linkage
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to compounds which are effective as catalysts for dismutating superoxide and, more particularly, the manganese or iron complexes of substituted, unsaturated heterocychc pentaazacyclopentadecane ligands which catalytically dismutate superoxide.
  • the enzyme superoxide dismutase catalyzes the conversion of superoxide into oxygen and hydrogen peroxide according to equation (1) (hereinafter referred to as dismutation).
  • Reactive oxygen metabolites derived from superoxide have been demonstrated to contribute to the tissue pathology in a number of inflammatory diseases and disorders, such as reperfusion injury to the ischemic myocardium, inflammatory bowel disease, rheumatoid arthritis, osteoarthritis, atherosclerosis, hypertension, metastasis, psoriasis, organ transplant rejections, radiation-induced injury, asthma, influenza, stroke, burns and trauma. See, for example, Simic, M. G., et al, Oxygen Radicals in Biology and Medicine, Basic Life Sciences, Vol. 49, Plenum Press, New York and London, 1988; Weiss ⁇ J. Cell.
  • ligands are described as a pentaazacyclopentadecane macrocycle with various substituents on the carbons ofthe macrocycle, or with cyclic or heterocychc structures attached to the carbons ofthe macrocycle.
  • These compounds have been shown to possess catalytic superoxide dismutating activity as well as anti-inflammatory activity and to prevent oxidative damage.
  • these compounds have been shown to possess analgesic activity in the rat-paw carrageenan hyperalgesia model, U.S. App. Ser. No. 09/057,831.
  • Two such described analgesic SOD mimic compounds are Compound A and Compound B:
  • these substituted, unsaturated, nitrogen-containing heterocychc compounds are up to ten times more potent as pharmaceutical agents for anti- inflammatory and analgesic compositions and are as good as, or often better than, the parent unsubstituted compounds in applications such as the treatment of endotoxin- induced refractory hypotension.
  • the compounds ofthe present invention demonstrate unanticipated improvements in characteristics important for pharmaceuticals over the previously described pentaazacyclopentadecane complexes with unsubstituted nitrogen- containing heterocychc moieties.
  • the present invention is directed to low molecular weight catalysts for the dismutation of superoxide radicals (SOD mimics) useful as therapeutic agents for inflammatory disease states and disorders in which superoxide anions are implicated.
  • SOD mimics ofthe present invention are manganese or iron complexes of nitrogen- containing fifteen-membered macrocyclic ligands which comprise a substituted, unsaturated, nitrogen-containing heterocychc moiety, most preferably those with cyclohexyl, hydroxyl alkyl thio, alkyl (2-thioacetic acid) ester, benzyloxy, methoxyarylthio, alkoxycarbonylarylthio, and aryl (2-thioacetic acid) ester substituents.
  • the nitrogen-containing heterocychc moiety is aromatic, more preferably a pyridino moiety.
  • the present invention is also directed to the pentaazacyclopentadecane macrocycles which comprise a substituted, unsaturated, nitrogen-containing heterocychc moiety which are precursor ligands of these complexes.
  • the present invention is also directed to methods of making the above SOD mimics, specifically, novel methods of modifying the substituents on the heterocychc moiety after chelation with the transition metal ion.
  • the present invention is also directed towards pharmaceutical compositions comprising the SOD mimics ofthe present invention in amounts sufficient for the treatment or prevention of disease states or disorders.
  • the present invention is directed to methods of using these catalysts to treat various disease states and disorders in which superoxide anions are implicated.
  • FIGURE 1 A chart of mean arterial pressure data from endotoxemic rats, Example 16. ⁇ Group received saline (control); D Group received LPS only; ⁇ Group received LPS and an infusion of 0.25 mg/kg/hr of Compound A at 1 hour; A Group received LPS and an infusion of 0.25 mg/kg/hr of Compound A at 5 hours.
  • FIGURE 2 A chart of mean arterial pressure data from endotoxemic rats, Example 16. ⁇ Group received saline (control); ⁇ Group received LPS only; ⁇ Group received LPS and an infusion of 0.075 mg/kg/hr of Compound 25 at 3 hours.
  • FIGURE 3 A chart of mean arterial pressure data from endotoxemic rats, Example 16. ⁇ Group received saline (control); ⁇ Group received LPS only; ⁇ Group received LPS and an infusion of 0.075 mg/kg/hr of Compound 31 at 3 hours.
  • FIGURE 4 A chart of paw volume change in the rat paw carrageenan model, Example 14. ⁇ Group received carrageenan injection only; • Group received an infusion of 6 mg/kg/hr of Compound A 15 minutes before carrageenan injection.
  • FIGURE 5 A chart of paw volume change in the rat paw carrageenan model, Example 14. ⁇ Group received carrageenan injection only; • Group received an infusion of 10 mg/kg/hr of Compound 13 15 minutes before carrageenan injection.
  • FIGURE 6 A chart of paw volume change in the rat paw carrageenan model, Example 14. ⁇ Group received carrageenan injection only; • Group received an infusion of 1 mg/kg/hr of Compound 14 15 minutes before carrageenan injection, ⁇ Group received an infusion of 10 mg/kg/hr of Compound 14 15 minutes before carrageenan injection.
  • FIGURE 7 A chart of paw volume change in the rat paw carrageenan model, Example 14. ⁇ Group received carrageenan injection only; • Group received an infusion of 10 mg/kg/hr of Compound 25 15 minutes before carrageenan injection.
  • FIGURE 8 A chart of paw volume change in the rat paw carrageenan model,
  • Example 14 ⁇ Group received carrageenan injection only; • Group received an infusion of 10 mg/kg/hr of Compound 31 15 minutes before carrageenan injection.
  • FIGURE 9 A chart depicting the protective effects of Compound A in a rat model of live E.coli induced shock.
  • Figure 9a depicts the protective effect of Compound A in preventing a fall in MAP.
  • Figure 9b depicts the protective effect of Compound A in preventing a fall in heart rate.
  • FIGURE 10 Depicts the molecular structure of Compound 25 ofthe present invention.
  • SOD mimic means a low-molecular-weight catalyst for the conversion of superoxide anions into hydrogen peroxide and molecular oxygen.
  • These catalysts consist of an organic ligand having a pentaazacyclopentadecane portion and a chelated transition metal ion, preferably manganese or iron.
  • the term may include catalysts containing short-chain polypeptides (under 15 amino acids), or macrocyclic structures derived from amino acids, as the organic ligand.
  • the term explicitly excludes a superoxide dismutase enzyme obtained from any natural sources.
  • precursor ligand means the organic ligand of a SOD mimic without the chelated transition metal cation and charge neutralizing anions.
  • substituted means that the described moiety has one or more substituents comprising at least 1 carbon or heteroatom, and further comprising 0 to 22 carbon atoms, more preferably from 1 to 15 carbon atoms, and comprising 0 to 22, more preferably from 0 to 15, heteroatoms selected from the group consisting of : O, S, N, P, Si, B, F, Cl, Br, or I. These atoms may be arranged in a number of configurations, creating substituent groups which are unsaturated, saturated, or aromatic.
  • substituents include branched or unbranched alkyl, alkenyl, or alkynyl, cyclic, heterocychc, aryl, heteroaryl, allyl, polycycloalkyl, polycycloaryl, polycycloheteroaryl, imines, aminoalkyl, hydroxyalkyl, hydroxyl, phenol, amine oxides, thioalkyl, carboalkoxyalkyl, carboxylic acids and their derivatives, keto, ether, aldehyde, amine, amide, nitrile, halo, thiol, sulfoxide, sulfone, sulfonic acid, sulfide, disulfide, phosphonic acid, phosphinic acid, acrylic acid, sulphonamides, amino acids, peptides, proteins, carbohydrates, nucleic acids, fatty acids, lipids, nitro, hydroxylamines, hydroxamic acids, thiocarbonyls,
  • alkyl alone or in combination, means a straight-chain or branched-chain alkyl radical containing from 1 to about 22 carbon atoms, preferably from about 1 to about 18 carbon atoms, and most preferably from about 1 to about 12 carbon atoms.
  • radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl and eicosyl.
  • alkenyl alone or in combination, means an alkyl radical having one or more double bonds.
  • alkenyl radicals include, but are not limited to, ethenyl, propenyl, 1-butenyl, cis-2-butenyl, trans-2-butenyl, iso-butylenyl, cis-2-pentenyl, trans-2-pentenyl, 3 -methyl- 1-butenyl, 2,3-dimethyl-2-butenyl, 1-pentenyl, 1-hexenyl, 1-octenyl, decenyl, dodecenyl, tetradecenyl, hexadecenyl, cis- and trans-9-octadecenyl, 1,3-pentadienyl, 2,4-pentadienyl, 2,3-pentadienyl, 1,3-hexadienyl, 2,4-hexadienyl
  • alkynyl alone or in combination, means an alkyl radical having one or more triple bonds.
  • alkynyl groups include, but are not limited to, ethynyl, propynyl (propargyl), 1-butynyl, 1-octynyl, 9-octadecynyl, 1,3-pentadiynyl, 2,4-pentadiynyl, 1,3-hexadiynyl, and 2,4-hexadiynyl.
  • cycloalkyl alone or in combination means a cycloalkyl radical containing from 3 to about 10, preferably from 3 to about 8, and most preferably from 3 to about 6, carbon atoms.
  • examples of such cycloalkyl radicals include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and perhydronaphthyl.
  • cycloalkylalkyl means an alkyl radical as defined above which is substituted by a cycloalkyl radical as defined above.
  • examples of cycloalkylalkyl radicals include, but are not limited to, cyclohexylmethyl, cyclopentylmethyl,
  • cycloalkylcycloalkyl means a cycloalkyl radical as defined above which is substituted by another cycloalkyl radical as defined above.
  • examples of cycloalkylcycloalkyl radicals include, but are not limited to, cyclohexylcyclopentyl and cyclohexylcyclohexyl.
  • cycloalkenyl alone or in combination, means a cycloalkyl radical having one or more double bonds.
  • examples of cycloalkenyl radicals include, but are not limited to, cyclopentenyl, cyclohexenyl, cyclooctenyl, cyclopentadienyl, cyclohexadienyl and cyclooctadienyl.
  • cycloalkenylalkyl means an alkyl radical as defined above which is substituted by a cycloalkenyl radical as defined above.
  • examples of cycloalkenylalkyl radicals include, but are not limited to, 2-cyclohexen-l-ylmethyl, 1-cyclopenten-l-ylmethyl, 2-(l -cyclohexen- l-y ⁇ )ethyl, 3-(l-cyclopenten-l-yl)propyl, 1 -( 1 -cyclohexen- 1 -ylmethyl)pentyl, 1 -( 1 -cyclopenten- 1 -yl)hexyl, 6-(l-cyclohexen-l-yl)hexyl, 1-(1 -cyclopenten- l-yl)nonyl and 1-(1 -cyclohexen- l-yl)nonyl.
  • alkylcycloalkyl and alkenylcycloalkyl mean a cycloalkyl radical as defined above which is substituted by an alkyl or alkenyl radical as defined above.
  • alkylcycloalkyl and alkenylcycloalkyl radicals include, but are not limited to, 2-ethylcyclobutyl, 1-methylcyclopentyl, 1-hexylcyclopentyl, 1-methylcyclohexyl, l-(9-octadecenyl)cyclopentyl and l-(9-octadecenyl)cyclohexyl.
  • alkylcycloalkenyl and “alkenylcycloalkenyl” means a cycloalkenyl radical as defined above which is substituted by an alkyl or alkenyl radical as defined above.
  • alkylcycloalkenyl and alkenylcycloalkenyl radicals include, but are not limited to, l-methyl-2-cyclopentyl, l-hexyl-2-cyclopentenyl, l-ethyl-2-cyclohexenyl, l-butyl-2-cyclohexenyl, l-(9-octadecenyl)-2-cyclohexenyl and 1 -(2-pentenyl)-2-cyclohexenyl.
  • aryl alone or in combination, means a phenyl or naphthyl radical which optionally carries one or more substituents selected from alkyl, cycloalkyl, cycloalkenyl, aryl, heterocycle, alkoxyaryl, alkaryl, alkoxy, halogen, hydroxy, amine, cyano, nitro, alkylthio, phenoxy, ether, trifluoromethyl and the like, such as phenyl, p-tolyl, 4-methoxyphenyl, 4-(tert-butoxy)phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-hydroxyphenyl, 1 -naphthyl, 2-naphthyl, and the like.
  • aralkyl alone or in combination, means an alkyl or cycloalkyl radical as defined above in which one hydrogen atom is replaced by an aryl radical as defined above, such as benzyl, 2-phenylethyl, and the like.
  • heterocychc means ring structures containing at least one other kind of atom, in addition to carbon, in the ring.
  • the most common ofthe other kinds of atoms include nitrogen, oxygen and sulfur.
  • heterocychcs include, but are not limited to, pyrrohdinyl, piperidyl, imidazolidinyl, tefrahydrofuryl, tefrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, triazolyl and tetrazolyl groups.
  • saturated, partially saturated or unsaturated cyclic means fused ring structures in which 2 carbons ofthe ring are also part ofthe fifteen-membered macrocychc ligand.
  • the ring structure can contain 3 to 20 carbon atoms, preferably 5 to 10 carbon atoms, and can also contain one or more other kinds of atoms in addition to carbon. The most common ofthe other kinds of atoms include nitrogen, oxygen and sulfur.
  • the ring structure can also contain more than one ring.
  • saturated, partially saturated or unsaturated ring structure means a ring structure in which one carbon ofthe ring is also part ofthe fifteen-membered macrocychc ligand.
  • the ring structure can contain 3 to 20, preferably 5 to 10, carbon atoms and can also contain nitrogen, oxygen and/or sulfur atoms.
  • nitrogen containing heterocycle means ring structures in which 2 carbons and a nitrogen ofthe ring are also part ofthe fifteen-membered macrocychc ligand.
  • the ring structure can contain 2 to 20, preferably 4 to 10, carbon atoms, can be substituted or unsubstituted, partially or fully unsaturated or saturated, and can also contain nitrogen, oxygen and/or sulfur atoms in the portion ofthe ring which is not also part ofthe fifteen-membered macrocychc ligand.
  • disease states and disorders in which superoxide anions are implicated means any disease state or disorder in which superoxide anions, or the products of reactions involving superoxide anions (such as peroxynifrate), are known or suspected to be a factor in the progression ofthe disease state or disorder.
  • diseases states and disorders are inflammation and ischemic reperfusion injury.
  • organic acid anion refers to carboxylic acid anions having from about 1 to about 18 carbon atoms.
  • halide means chloride, floride, iodide, or bromide.
  • R groups means all ofthe R groups attached to the carbon atoms ofthe macrocycle, i.e., R, R', Rl, R'l, R2, R'2, R3, R'3, R4, R'4, R5, R'5, R6, R'6, R7, R'7, R8, R'8, R9.
  • the present invention is directed to nitrogen-containing fifteen-membered macrocychc ligands, and their complexes with transition metals, which are described by the formula:
  • a nitrogen ofthe macrocycle and the two adjacent carbon atoms to which it is attached independently form a substituted, unsaturated, nitrogen containing heterocycle W having 2 to 20 carbon atoms, more preferably 4 to 10 carbon atoms, which may be an aromatic heterocycle wherein the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part ofthe heterocycle and the macrocycle are absent; and wherein R, R perpetrat R 2 , R' 2 , R 3 , R' 3 , R 4 , R' 4 , R ⁇ R' 5 , R 6 , R' 6 , R 7 , R' 7 , R 8 , R' g , R,, and R' 9 independently represent hydrogen, or substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylcycloalkyl,
  • X, Y, and Z may be independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide
  • the SOD mimics ofthe present invention can have any combinations of substituted or unsubstituted R groups, saturated, partially saturated or unsaturated cyclics, ring structures, nitrogen containing heterocycles, or straps as defined above.
  • the "R” groups attached to the carbon atoms ofthe macrocycle can be in the axial or equatorial position relative to the macrocycle.
  • the "R” group is other than hydrogen or when two adjacent “R” groups, i.e., on adjacent carbon atoms, together with the carbon atoms to which they are attached form a saturated, partially saturated or unsaturated cyclic or a nitrogen containing heterocycle, or when two R groups on the same carbon atom together with the carbon atom to which they are attached form a saturated, partially saturated or unsaturated ring structure, it is preferred that at least some ofthe "R” groups are in the equatorial position for reasons of improved activity and stability. This is particularly true when the complex contains more than one "R" group which is not hydrogen.
  • Preferred compounds ofthe present invention are those described by the following formula:
  • R groups, W, M, X, Y, and Z are as defined above, and wherein U and V are saturated cyclic structures, containing between 3 and 20, preferably between 4 and 10 carbon atoms and forming a cycloalkyl ring with the carbon atoms to which they are attached.
  • U and V are two trans- cyclohexano fused rings.
  • W is a substituted pyridine, and R, R perpetrat and the H on the nitrogen ofthe macrocycle within W are absent.
  • W is a substituted pyridine, and U and V are trans-cyclohexano fused rings.
  • Preferred substituents on W are those which increase the potency ofthe catalyst for pharmaceutical applications.
  • lipophilic substituents are preferred when the target ofthe catalyst is a hydrophobic tissue ofthe patient.
  • certain substituents generally increase the potency ofthe catalyst for use in pharmaceutical compositions. These preferred substituents include cyclohexyl, hydroxyl alkyl thio, alkyl
  • (2-thioacetic acid) esters 2,3-thioacetic acid esters, benzyloxy, methoxyarylthio, alkoxycarbonylarylthio, and aryl
  • the catalyst will function with a k cat of about 10 "6 to 10 "8 .
  • the complexes derived from Mn(II) and the general class of C-substituted [15]aneN 5 ligands described herein have been characterized using cyclic voltammetry to measure their redox potential.
  • the C-substituted complexes described herein have reversible oxidations of about +0.7 v (SHE). Coulometry shows that this oxidation is a one-electron process; namely it is the oxidation ofthe Mn(II) complex to the Mn(III) complex.
  • SOD catalysts the Mn(III) oxidation state is involved in the catalytic cycle.
  • Mn(III) complexes of all these ligands are equally competent as SOD catalysts, since it does not matter which form (Mn(II) or Mn(III)) is present when superoxide is present because superoxide will simply reduce Mn(III) to Mn(II), liberating oxygen.
  • the mechanism described in Riley, et al., 1999 is a reasonable approximation of how these catalysts dismutate superoxide.
  • the ligand In order for the complex to exhibit superoxide dismutase activity, the ligand should be able to fold into a conformation that allows the stabilization of an octahedral complex between an axial ligand and the five nitrogens ofthe ligand ring. If a compound contains several conjugated double bonds within the main 15-membered ring ofthe ligand, which hold the ring in a rigid conformation, the compound would not be expected to exhibit catalytic activity.
  • R groups attached to the macrocychc ligand which lock it in a planar conformation would be expected to be poor catalysts.
  • One of ordinary skill in the art would not be surprised that these types of derivatives would lack superoxide dismutase activity Specifically, one of skill in the art would avoid materially changing the flexibility ofthe macrocycle by adding many large groups which would cause steric hindrance, or placing too many double bonds into the main ring. This effect would also be present in certain geometric arrangements of smaller R groups which constrain the complex to a rigid, planar geometry. Given these examples and guidelines, one of ordinary skill would be able to recognize which ofthe described pentaazacyclopentadecane complexes ofthe present invention would retain superoxide dismutating activity.
  • the catalysts ofthe present invention may be produced by the methods disclosed in U.S. Pat. No. 5,610,293. However, it is preferred that the catalysts of the present invention be synthesized by the template method, diagrammed below. This synthesis method is advantageous over previously disclosed methods in that cyclization yields utilizing the template method are usually about 90%, as compared to about 20% with previous methods.
  • Several diamines are commercially available as starting materials, or a diamine may be synthesized. The diamines are reacted with titryl chloride in anhydrous methylene chloride at 0°C and allowed to warm to room temperature overnight, with stirring. The product is then combined with glyoxal in methanol and stirced for 16 hours.
  • the glyoxal bisimine product is then reduced with a borohydride in THF. If a symmetrical product is desired, one diamine may be used as the starting material. In addition, a substituted glyoxal may be used if groups pendant from the macrocycle opposite the pyridine are desired (R 5 and Rg) Commercially available tetraamines may also be used in place ofthe reduced glyoxal bisimine. After reduction ofthe glyoxal bisimine, the product is combined with a 2,6 dicarbonyl substituted pyridine, such as 2,6-dicarboxaldehyde pyridine or 2,6 diacetylpyridine, and a salt of manganese or iron under basic conditions.
  • a 2,6 dicarbonyl substituted pyridine such as 2,6-dicarboxaldehyde pyridine or 2,6 diacetylpyridine
  • the transition metal ion serves as a template to promote cyclization ofthe substituted pyridine and the tefraamine.
  • 2,6 dicarbonyl substituted pyridines are available commercially, allowing for the facile production of a variety of ligands with groups pendant from the macrocycle proximal to the pyridine (R 2 and R ⁇ ,).
  • Pyridines with one or more substituents (R A , R B and R ⁇ .) are used when synthesizing the catalysts ofthe present invention in order to obtain the substituted, unsaturated heterocychc moiety. After cyclization, the product is reduced with ammonium formate and a palladium catalyst over a period of 3-4 hours.
  • the bisimine may be reduced with a hydride reductant such as NaBH 4 , or with hydrogen gas and a metal catalyst.
  • a hydride reductant such as NaBH 4
  • hydrogen gas and a metal catalyst may also be substituted at the same carbons.
  • the bisimine produced in the template cyclization reaction step above may be reduced by more conventional means using hydrogen gas, it is preferred that the bisimine be reduced with ammonium formate in the presence of a catalyst, as illustrated in Example 2.
  • the prefened catalyst for use in this process comprises palladium, although a catalyst comprising other catalytic metals such as nickel, rhodium, platinum, platinum oxide, and ruthenium would also be potentially suitable.
  • This process offers the advantages of increased safety and high reduction efficiency ofthe imine bonds, while preserving the double bonds ofthe pyridine groups in the heterocychc moiety ofthe prefened compounds.
  • this method can be accomplished in a more concentrated medium as compared to hydrogen or borohydride reduction, allowing for faster reaction times.
  • Example 7 Several advantages are realized by using this process ofthe present invention. First, one is able to use commercially available or relatively easily synthesized reactants in the above template cyclization synthesis, such as 4-chloro-2,6 dicarboxaldyhyde pyridine, and then modify the resultant chelated macrocychc ligand without side reactions with the substituted group. Second, because this method allows modification ofthe chelated ligand, no post-modification reaction with manganese chloride is necessary, simplifying the synthesis process. A leaving-group-substituted pyridine pentaazacyclopentadecane chelated ligand is used as the starting material in the modification reaction.
  • reactants in the above template cyclization synthesis such as 4-chloro-2,6 dicarboxaldyhyde pyridine
  • nucleophilic 4- pyridino substituents which are good leaving groups are the halides. Cl, Br, and I are more preferred substituents.
  • a nucleophile (1 eq.), dropwise, and the reaction mixture stirred overnight. The solvent is then removed, in vacuo, the resulting mixture extracted with methylene chloride, and then concentrated down, in vacuo.
  • the SOD mimic catalyst may then be purified by flash column chromatography.
  • Nucleophiles for use in this modification reaction may be any strong nucleophile. Applicants have found that thiolates have provided a wide array of post-chelation modification reagents useful for making the compounds ofthe present invention.
  • manganese is usually used as the chelated transition metal ion in the examples of this disclosure, it is to be understood that the disclosed ligands may be just as easily complexed with iron (II) or iron (III) cations obtained from salts such as FeCl 3 . In general, better catalytic activity has been observed with the use of manganese as the chelated transition metal ion, although k cat 's which are as high as 10 "7 are still observed with the use of iron. Thus, manganese is preferred as the chelated transition metal ion in the complexes ofthe present invention.
  • the pentaazamacrocycles ofthe present invention can possess one or more asymmetric carbon atoms and are thus capable of existing in the form of optical isomers as well as in the form of racemic or nonracemic mixtures thereof.
  • the optical isomers can be obtained by resolution ofthe racemic mixtures according to conventional processes, for example by formation of diastereoisomeric salts by treatment with an optically active acid.
  • appropriate acids are tartaric, diacetyltartaric, dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid and then separation ofthe mixture of diastereoisomers by crystallization followed by liberation ofthe optically active bases from these salts.
  • a different process for separation of optical isomers involves the use of a chiral chromatography column optimally chosen to maximize the separation ofthe enantiomers.
  • Still another available method involves synthesis of covalent diastereoisomeric molecules by reacting one or more secondary amine group(s) ofthe compounds ofthe invention with an optically pure acid in an activated form or an optically pure isocyanate.
  • the synthesized diastereoisomers can be separated by conventional means such as chromatography, distillation, crystallization or sublimation, and then hydrolyzed to deliver the enantiomerically pure ligand.
  • the optically active compounds of the invention can likewise be obtained by utilizing optically active starting materials, such as natural amino acids.
  • the compounds ofthe present invention have been shown to have remarkable potency and utility in several models of disease.
  • Example 14 the utility ofthe present compounds for the treatment of pain and inflammation is demonstrated in the rat paw carrageenan model.
  • the substituted, unsaturated, nitrogen-containing heterocychc compounds sometimes differ remarkably from the base compound (compound A) in terms of their potency, onset of analgesia, and duration of effect.
  • the substitution of various small ester groups on the nitrogen containing heterocyclo moiety produces a very rapid onset of analgesic action.
  • the benzyl ether substituted complex is particularly potent in this model as compared to Compound A or B, both of which have higher rates of catalytic superoxide dismutation.
  • Example 15 the efficacy ofthe compounds in a murine model of opioid tolerance reversal is shown by both iv and subcutaneous administration.
  • the compounds of the present invention show astonishing efficacy in the prevention of morphine tolerance in this model. For instance, in iv administration, Compounds 13 and 14 show a very significant reversal of morphine tolerance at 1/30 ofthe concentration necessary to achieve roughly 1/2 the effect with Compound A.
  • Compound 28 is shows a 100% reversal of morphine tolerance at 1/100 the concentration needed for the same effect with Compound A, in iv administration. Similar results were obtained with subcutaneous administration, in which Compound 3 showed 100% reversal of morphine tolerance at 1/100 the dose necessary to achieve the same effect with Compound A. Thus, the compounds ofthe present invention show remarkable utility for preventing or reversing opioid tolerance.
  • Compounds 25 and 31 were also tested for their ability to prevent refractory hypotension in an endotoxemic rat model, Example 16. Both of these compounds were effective at preventing hypotension in endotoxemic animals at 1/3 the dose used to achieve a similar effect with Compound A.
  • the compounds or complexes ofthe present invention are can be utilized to treat numerous disease states and disorders in a patient in need thereof.
  • patient and “subject” includes human and non-human animals in need of treatment.
  • disease states and disorders include, but are not limited to: reperfusion injury to an ischemic organ, such as reperfusion injury to the ischemic myocardium, general inflammation, inflammatory bowel disease, rheumatoid arthritis, osteoarthritis, hypertension, psoriasis, organ transplant rejections, refractory hypotension, organ preservation, radiation-induced injury, platelet aggregation, stroke, autoimmune diseases, adult respiratory distress, carcinogenesis, severe chronic pain, hyperalgesia, and sepsis.
  • reperfusion injury to an ischemic organ such as reperfusion injury to the ischemic myocardium, general inflammation, inflammatory bowel disease, rheumatoid arthritis, osteoarthritis, hypertension, psoriasis, organ transplant rejections, refractory
  • the complexes of this invention are excellent analgesics and can be used to treat or prevent pain in a subject arising from any hyperalgesic state.
  • the complexes further have activity to prevent or reduce tolerance to opiates, and to potentiate the analgesic activity of opiates without potentiating the respiratory depression associated with opiates.
  • the complexes are useful in treating withdrawal symptoms associated with addiction to opiates, nicotine, or other drugs.
  • the complexes of this invention can also be used systemically or topically to prevent or reverse free oxygen radical-mediated symptoms of aging, such as skin wrinkling, and to prevent or reverse environmental damage caused by exposure to ultraviolet radiation or chemical agents.
  • Total daily dose administered to a subject in single or divided doses may be in amounts, for example, from about 0.00025 to about 20 mg/kg body weight daily, more preferably from about 0.001 to about 10 mg/kg body weight daily, and more usually about 0.01 to about 3 mg/kg body weight daily, when given as a parenteral injection or continuous infusion.
  • Dosage unit compositions may contain such amounts of sub- multiples thereof to make up the daily dose.
  • the amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the subject treated and the particular mode of administration. For instance, systems such as transdermal administration or oral administration, which are substantially less efficient delivery systems, may require dosages at least an order of magnitude above those required for parenteral administration.
  • the dosage regimen for treating a disease condition with the compounds and/or compositions of this invention is selected in accordance with a variety of factors, including the type, age, weight, sex, diet and medical condition ofthe patient, the severity ofthe disease, the route of administration, pharmacological considerations such as the activity, efficacy, pharmokinetic and toxicology profiles ofthe particular compound employed, whether a drug delivery system is utihzed and whether the compound is administered as part of a drug combination.
  • the dosage regimen actually employed may vary widely and therefore may deviate from the preferred dosage regimen set forth above.
  • Those of ordinary skill in the art can readily determine appropriate dosages for any particular subject based on the teachings in this specification and routine analysis ofthe subject.
  • the compounds ofthe present invention may be administered by any technique known to those of ordinary skill, including but not limited to, orally, parenterally, by inhalation spray, rectally, topically or by nasal, vaginal or ocular administration, in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. Topical administration may also involve the use of transdermal administration such as transdermal patches or iontophoresis devices.
  • parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, intrathecal or infusion techniques.
  • sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
  • acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid find use in the preparation of injectables.
  • Suppositories for rectal administration ofthe drug can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter and polyethylene glycols which are solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug.
  • Solid dosage forms for oral administration may include capsules, tablets, pills, powders, granules and gels.
  • the active compound may be admixed with at least one inert diluent such as sucrose, lactose, or starch.
  • Such dosage forms may also comprise, as in normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate.
  • the dosage forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings.
  • Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.
  • While the compounds ofthe invention can be administered as the sole active pharmaceutical agent, they can also be used in combination with one or more compounds which are known to be effective against the specific disease state that one is targeting for treatment.
  • the compounds ofthe present invention make exceptional catalysts for the dismutation of superoxide. Thus, they can be used in this catalytic capacity in a variety of in vivo and in vitro applications where a reduction in superoxide concentration is desired.
  • the SOD mimic compounds ofthe present invention can also be added to rinse or storage solutions for organs and tissues, such as for organ transplantation or for surgical rinses.
  • excised organs are often placed in a preservation solution prior to transplant into a recipient.
  • Inclusion of at least one species of SOD mimic in a preservation solution usually at a concentration of about 0.01 mM to 10 mM, is desirable for reducing damage due to ischemia during storage and reperfusion injury following reimplantation in the recipient.
  • Various solutions described in the art are suitable for the inclusion of these compounds ofthe invention, including but not limited to those described in U.S. Pat. No. 5,145,771; Beyersdorf (1990) Chem Abst. 113: 84849w; U.S. Pat. No.
  • the compounds ofthe present invention can also be added to extravasated blood for transfusion to inhibit oxyradical damage to the blood cells and components during storage; similarly, these compounds can also reduce oxyradical damage to blood cells in vivo.
  • the SOD mimic ofthe present invention is present in the rinse or storage solution at a concentration of about 0.001 mM to about 10 mM, and most usually is present at 1 mM.
  • a suitable rinse solution comprises Ringer's solution (102 mM NaCl, 4 mM KCl, 3 mM CaC12, 28mM sodium lactate, pH 7.0) or Ringer's solution with 0.1 mM adenosine, and Compound 1 at a final concentration of 1 mM.
  • the rinse solution can further comprise additional antioxidants
  • Preservation or rinse solutions containing a SOD mimic of the present invention can be used to provide enhanced storage or irrigation of organs (e.g., kidney, liver, pancreas, lung, fetal neural tissue, heart, vascular grafts, bone, ligament, tendon, skin)which is believed to enhance the viability ofthe tissue and increase resistance to oxidative damage (e.g., as a consequence of ischemia/reperfusion).
  • organs e.g., kidney, liver, pancreas, lung, fetal neural tissue, heart, vascular grafts, bone, ligament, tendon, skin
  • the capacity ofthe compounds ofthe present invention to catalyze the decomposition of reactive oxygen species can be used to advantage to inhibit or slow damage to biological tissues and cells.
  • oxyradical-induced damage to connective tissues e.g., collagen
  • SOD mimics ofthe present invention can also be formulated into a lipophilic base (or, if desired, an aqueous carrier) for topical application in cosmetics or sunburn-prevention creams and lotions.
  • a typical cosmetic or sunburn-prevention cream or lotion will comprise about between 1 mg to 50 mg of SOD mimic compound per gram of cosmetic or sunburn-prevention cream or lotion.
  • the compounds ofthe present invention can be formulated into a cosmetic base for topical application and/or for reducing oxidation ofthe cosmetic by molecular oxygen and oxyradicals.
  • the pharmaceutical/cosmetic compositions ofthe present invention formulated as solutions typically include a pharmaceutically- or cosmetically-acceptable organic solvent.
  • pharmaceutically-acceptable organic solvent and “cosmetically-acceptable organic solvent” refer to an organic solvent which, in addition to being capable of having dispersed or dissolved therein the salen-metal compound, and optionally also an anti-inflammatory agent, also possesses acceptable safety (e.g.
  • a solvent is isopropanol.
  • suitable organic solvents include: propylene glycol, polyethylene glycol (200-600), polypropylene glycol
  • emollients refer to materials used for the prevention or relief of dryness, as well as for the protection ofthe skin.
  • suitable emollients are known and may be used herein. Sagarin, Cosmetics, Science and Technology, 2nd Edition, Vol. 1, pp. 32-43 (1972), incorporated herein by reference, contains numerous examples of suitable materials.
  • Particularly useful emollients which provide skin conditioning are glycerol, hexanetriol, butanetriol, lactic acid and its salts, urea, pyrrolidone carboxylic acid and its salts, amino acids, guanidine, diglycerol and triglycerol.
  • Preferred skin conditioning agents are the propoxylated glycerol derivatives.
  • the invention also provides methods for preventing food spoilage and oxidation by applying to foodstuffs an effective amount of at least SOD mimic compound ofthe present invention, optionally in combination with at least one additional food preservative agent
  • the invention relates to antioxidant compositions and methods of use in inhibiting formation of undesired hydrocarbon polymers generated via free radical-mediated polymerization mechanisms, especially oxyradical-mediated polymerization and/or oxyradical-mediated rancidification or gum formation.
  • the SOD mimic compounds ofthe invention can be applied to a variety of hydrocarbons to reduce undesired oxidation and/or polymerization, or to quench a polymerization reaction at a desired state of polymer formation (e.g., at a desired average chain length).
  • saturated and unsaturated hydrocarbons include: petroleum distillates and petrochemicals, turpentine, paint, synthetic and natural rubber, vegetable oils and waxes, animal fats, polymerizable resins, polyolefin, and the like.
  • the compounds ofthe present invention may also be used to protect cells and tissues from free radical-producing agents, such as ionizing radiation and chemotherapeutic agents (e.g., bleomycin).
  • a protective dosage comprising at least about 0.001 mg of SOD mimic/kg body weight is administered by one or more of several routes (e.g., oral, intravenous, intraperitoneal, intragastric lavage, enema, portal vein infusion, topical, or inhalation of mist) to protect normal cells, for example, against free radical toxicity associated with chemotherapy or radiotherapy of a neoplasm.
  • the compounds ofthe present invention are preferably pre-administered to the patient prior to the commencement ofthe chemotherapy and/ or radiotherapy, usually within about 24 hours of commencement, and preferably within about 3-6 hours of commencement ofthe chemotherapy and/ or radiotherapy.
  • the compounds may be continually administered to the patient during the course of therapy.
  • the SOD mimics ofthe present invention also can be administered to individuals to prevent radiation injury or chemical injury by free radical generating agents. Military personnel and persons working in the nuclear, nuclear medicine, and/or chemical industries may be administered the compounds ofthe present invention prophylactically.
  • chemoprotective agents may also be used as chemoprotective agents to prevent chemical carcinogenesis; particularly by carcinogens which form reactive epoxide intermediates (e.g., benzo->a!-pyrene, benzanthracene) and by carcinogens or promoting agents which form free radicals directly or indirectly (e.g., phenobarbital, TPA, benzoyl peroxiprolieroxisome prohferators: ciprofibrate, clofibrate). Persons exposed to such chemical carcinogens are pretreated with the compounds ofthe present invention to reduce the incidence or risk of developing neoplasia.
  • carcinogens which form reactive epoxide intermediates
  • N,N'-Bis ⁇ (lR,2R)-[2-(Triphenylmethylamino)]cyclohexyl ⁇ - 1 ,2-diaminoethane N,N'-Bis ⁇ (lR,2R)-[2-(Triphenylmethylamino)]cyclohexyl ⁇ - 1 ,2-diaminoethane.
  • the glyoxal bisimine of N-(triphenylmethyl)-(lR,2R)-diaminocyclohexane (586 g,
  • N,N'-Bis ⁇ (lR,2R)-[2-(amino)]cyclohexyl ⁇ -l,2-diaminoethane tetrahydrochloride To a solution of N,N'-bis ⁇ (lR,2R)-[2-(triphenylmethylamino)]cyclohexyl ⁇ -l,2- diaminoethane (590 g, 798 mmol) in acetone (3 L) was added concentrated HC1 (1.5 L). The reaction was stirred for 2 hours and concentrated. The residue was partitioned between water (2 L) and CH 2 C1 2 (1 L).
  • the purified bisimine precursor to Compound B (15.0 g, 29.6 mmol) was dissolved in 1.5 L of anhydrous MeOH and the flask flushed with nitrogen for a few minutes, then 3% P ⁇ VC (7.5 g, 50% by weight) was added. As the suspension was heated, solid ammonium formate (7.5 g, 118.9 mmol, 4 equiv.) was carefully added. One hour after reflux was attained, a second portion of formate (3.75 g, 59.5 mmol, 2 equiv.) was added.
  • the black suspension was allowed to cool to RT after 2.5 h of reflux (at this point the supernatant was virtually colorless), and filtered through a 1 /2-inch bed of alumina (Al 2 O 3 ,
  • Water for buffer solutions was delivered from an in-house deionized water system to a Barnstead Nanopure Ultrapure Series 550 water system and then double distilled, first from alkaline potassium permanganate and then from a dilute EDTA solution.
  • a solution containing 1.0 g of potassium permanganate, 2 liters of water and additional sodium hydroxide necessary to bring the pH to 9.0 were added to a 2-liter flask fitted with a solvent distillation head. This distillation will oxidize any trace of organic compounds in the water.
  • the final distillation was carried out under nitrogen in a 2.5-liter flask containing 1500 ml of water from the first still and 1.0 x 10 "6 M EDTA.
  • This step will remove remaining trace metals from the ultrapure water.
  • the 40-cm vertical arm was packed with glass beads and wrapped with insulation. This system produces deoxygenated water that can be measured to have a conductivity of less than 2.0 nanoohms/cm2.
  • the stopped-flow spectrometer system was designed and manufactured by
  • Aqueous solutions to be mixed with the DMSO solution of superoxide were prepared using 80 mM concentrations ofthe Hepes buffer, pH 8.1 (free acid+Na form).
  • One ofthe reservoir syringes was filled with 5 ml ofthe DMSO solution while the other was filled with 5 ml ofthe aqueous buffer solution.
  • the entire injection block, mixer, and spectrometer cell were immersed in a thermostated circulating water bath with a temperature of 21.0° ⁇ 0.5° C.
  • a baseline average was obtained by injecting several shots ofthe buffer and DMSO solutions into the mixing chamber. These shots were averaged and stored as the baseline.
  • the first shots to be collected during a series of runs were with aqueous solutions that did not contain catalyst. This assures that each series of trials were free of contamination capable of generating first-order superoxide decay profiles. If the decays observed for several shots ofthe buffer solution were second-order, solutions of manganese(II) complexes could be utilized.
  • the potential SOD catalyst was screened over a wide range of concentrations. Since the initial concentration of superoxide upon mixing the DMSO with the aqueous buffer was about 1.2 times 10-4 M, we wanted to use a manganese (II) complex concentration that was at least 20 times less than the substrate superoxide. Consequently, we generally screened compounds for superoxide dismutating activity using concentrations ranging from 5 x 10-7 to 8 x
  • Catalytic rate constants for dismutation of superoxide by manganese(II) complexes were determined from linear plots of observed rate constants (kobs) versus the concentration ofthe manganese(II) complexes, kobs values were obtained from linear plots of In absorbance at 245 nm versus time for the dismutation of superoxide by the manganese(II) complexes.
  • Example 1 (2.80 g, 7.00 mmol) in absolute ethanol (70 mL) was added KOH (1.79 g - 88 %, 28.0 mmol) and the mixture was stined at room temperature under an argon atmosphere. After 30 minutes, MnCl 2 (881 mg, 7.00 mmol) was added and the suspension was stirred for an additional 30 minutes. 4-Cyclohexyl-2,6- pyridinedicarboxaldehyde (1.52 g, 7.00 mmol) was then added to the dark green mixture which was refluxed.
  • the flask was allowed to reach room temperature and left at room temperature for 16 h.
  • the flask contained a white precipitate which was filtered and washed with chloroform (30 mL).
  • the white solid was dissolved in hot THF (300 mL), dried over magnesium sulfate and filtered, then concentrated in vacuo to afford 32.1 g 4-chloro-2,6-dihydroxymethyl pyridine as a white sohd.
  • the filtrate was concentrated.
  • the resulting white solid was heated in THF (500 mL), dried over magnesium sulfate and filtered.
  • Oxalyl chloride (126.93 g, 154 mmol) and CH 2 C1 2 (80 mL) were placed in a 1 L, 3-neck round bottomed flask. The solution was cooled to -60 °C. To the cooled solution, DMSO (24 mL) in CH 2 C1 2 (80 mL) was added over a 5 minutes period via dropping funnel. After 10 minutes, 4-chloro-2,6-dihydroxymethyl pyridine (12.13 g, 69.9 mmol) in DMSO (40 mL) was added over 5 minutes, also via dropping funnel.
  • Example 6 (1.00 g, 1.94 mmol) was placed in a flask and dissolved in DMF (80 mL). In a separate flask, 2-diethylaminoethanethiol » HCl (364 mg, 2.14 mmol) was dissolved in DMF (20 mL). The flask was cooled to 0°C in an ice- water bath. To the flask was added ⁇ aH (102 mg, 8.5 mmol). After stirring for 30 minutes, the 2- diethylaminoethanethiolate solution was added to the
  • Manganese( ⁇ )dichloro(4R,9R,14R,19R-24-chloro-3,10,13,20,26- pentaazatetracyclo-[20.3.1.0 4,9 .0 14,19 ]hexacosa-l(26),22(23),24-triene) prepared as in Example 6 (1.01 g, 1.96 mmol) was placed in a flask and dissolved in DMF (60 mL). In a separate flask, 2-mercaptopropane (165 mg, 2.15mmol) was dissolved in DMF (60 mL). The flask was cooled to 0°C in an ice-water bath.
  • the concentrate was extracted with saturated ⁇ aCl solution (250 mL) and water (250 mL) then extracted with CH 2 C1 2 (250 mL). The aqueous layer was washed with several portions of CH 2 C1 2 . The organic fractions were combined, dried over ⁇ a 2 SO 4 and concentrated in vacuo.
  • the crude material was purified by silica gel chromatography (Aldrich 200-400 mesh, 60 A). The product was eluted through the column with 1% MeOH/CH 2 Cl 2 increasing slowly to
  • the concentrate was extracted with saturated NaCl solution (250 mL), water (250 mL) and CH 2 C1 2 (250 mL). The aqueous layer was washed with several portions of CH 2 C1 2 . The organic fractions were combined, dried over Na 2 SO 4 and concentrated in vacuo.
  • the crude material was purified by silica gel chromatography (Aldrich 200-400 mesh, 60 A). The product was eluted through the column with 1 % MeOH/CH 2 Cl 2 increasing slowly to 2% MeOH/CH 2 Cl 2 .
  • Example 6 (1.00 g, 1.93 mmol) was placed in a flask and dissolved in DMF (80 mL). In a separate flask, cyclohexyhnercaptan (247 mg, 2.12 mmol) was dissolved in DMF (20 mL). The flask was cooled to 0°C in an ice-water bath. To the flask was added NaH (51 mg, 2.13 mmol). After stirring for 30 minutes, the cyclohexylthiolate solution was added to the
  • Sodium hydride (3.40 g, 142 mmol) was weighed into a 500 mL flask equipped with a stirbar under inert atmosphere. Anhydrous DMF (230 mL) was added to the sodium hydride and a slurry was created. The flask was cooled in a ice- water bath and ethyl thioglycolate (16.97 mL, 155 mmol) was gradually added to the slurry. After gas evolution ceased, the ice bath was removed and 120 mL ofthe thiolate solution was added to the solution of manganese complex. The cooling bath was removed. After 4.6 hours, an additional 100 mL of thiolate solution was added to the reaction.
  • Methylthioglycolate (190 ⁇ L, 2.12 mmol) was added to a slurry of NaH (50.9 mg, 2.12 mmol) in 10 mL of anhydrous DMF which was cooled in an ice- water bath.
  • the mixture was allowed to warm to room temperature.
  • step D The orange-red ethanol solution from step D was diluted with water (200 mL). Palladium black (5 g) and ammonium formate (10 g) were added and the reaction was refluxed for 3 hours. The reaction was cooled to room temperature, filtered through celite, and the cake washed with water (500 mL) and ethanol (500 mL). The filtrate was evaporated to afford 10 g of Manganese(II)dichloro-
  • Oil in water emulsion as percentages by weight.
  • Rats Male Sprague-Dawley rats (175-200 g, Harlan Sprague Dawley, Indianapolis, IN, USA) were housed and cared for in accordance with the guidelines ofthe Institutional Animal Care and Use Committee and in accordance with NTH guidelines on laboratory animal welfare. Rats received a subplantar injection of ca ⁇ ageenan (0.1 ml of a 1% suspension in 0.85% saline) into the right hind paw. Paw volume was measured using plethysmometer (Ugo-Basile, Varese, Italy) immediately prior to the injection of ca ⁇ ageenan and thereafter at hourly intervals for up to 6 h.
  • ca ⁇ ageenan 0.1 ml of a 1% suspension in 0.85% saline
  • Edema was expressed as the increase in paw volume (ml) after ca ⁇ ageenan injection relative to the pre-injection value for each animal.
  • Drugs were administered intravenously (iv) in a volume of 2.5 ml/kg, 30 min prior or at least 3 h post ca ⁇ ageenan injection.
  • a hyperalgesic response to heat was determined in the same animals by Hargreaves method (Hargreaves et al, 1988). Rats were individually confined and acclimated to plexiglass chambers for 30 min.
  • a mobile unit consisting of a high intensity projector bulb was positioned to deliver a thermal stimulus directly to an individual hind paw from beneath the chamber.
  • the withdrawal latency period of injected and contralateral paws was determined to the nearest 0.1 sec with an electronic clock circuit and thermocouple. If the animal failed to respond by 20 sec the test was terminated. Each point will represent the change in withdrawal latency compared with control measurements taken prior to ca ⁇ ageenan injection.
  • the intraplantar injection of ca ⁇ ageenan in rats resulted in a time-dependent increase in paw volume and hyperalgesia that was maximal after 3-6h. As shown in
  • mice Male CD-I mice (Charles River, 28-35 gm) were allowed to feed ad libitum. Mice were housed 5-7 per cage in a temperature-controlled room with a 12-hr light-dark cycle. Nociceptive thresholds were measured by comparing hind paw escape latencies on a hot plate (Model 35, IITC Inc., Woodland Hills, CA) maintained at 57°C. Mice were placed on the heated surface enclosed by a transparent glass cylinder 25 cm high and 15 cm in diameter. Latency response was reported as the time to intermittent lifting or licking of the hind paws. A cut off latency of 20 sec was employed to prevent tissue damage in non- responsive animals. Determination of antinociception was assessed between 7:00 and 10:00 AM.
  • mice were rendered tolerant by twice daily subcutaneous injections of morphine (2 x 10 mg/kg day) for a four day period as evidenced by a decreased antinociceptive response to a 3 mg/kg challenge dose to morphine on day 5.
  • the latency to 3.0 mg/kg mo hine in naive mice ranged from 11-13 sec. 50 min. post injection and was assigned a maximal antinociceptive score of 100%.
  • Morphine was obtained from Mallinckrodt (St. Louis). Drugs were dissolved in saline except for the SOD mimic which was dissolved in sodium bicarbonate (26 mM, pH 8.3). Injection volumes were 0.01 mL/gm body weight.
  • mice The latency for naive mice (5.4 ⁇ 0.7 sec) was denoted as 0% analgesia.
  • Tolerant mice exhibited latencies of 6.8 ⁇ 0.7 sec after 3 mg/kg morphine as measured on day 5 (time when tolerance was observed).
  • the SOD mimics (mg/kg) were injected 5 min (iv studies) or 40 min (sc studies) prior to morphine and antinociception was measured 50 min later. 9-20 mice were used at each dose.
  • EXAMPLE 16 USE OF THE SOD MIMICS OF THE INVENTION IN THE TREATMENT OF REFRACTORY HYPOTENSION IN ENDOTOXEMIC RATS Male Sprague-Dawley rats (175-200 g, Harlan Sprague Dawley, Indianapolis, IN,
  • Rats were housed and cared for in accordance with the guidelines ofthe Institutional Animal Care and Use Committee and in accordance with NTH guidelines on laboratory animal welfare. Rats were anesthetized with inactin (100 mg/kg intraperitoneally). The trachea was cannulated to facilitate respiration and body temperature was maintained at 37°C by means of a heating pad for the entire duration ofthe experiment (9 hours). The left femoral vein was cannulated for the administration of drugs. After a 30 min stabilization period, lipopolysaccharide from E.
  • LPS lipoprotein
  • serotype 0111 :B4 was administered as a bolus intravenous (iv) injection at a volume of 0.3 ml and mean arterial pressure/heart rate monitored for 9 hours.
  • Control animals received isotonic saline at the same volume and by the same route. 1 , 3, or 5 hours after LPS, SOD mimic or vehicle (26 mM sodium bicarbonate buffer, pH 8. ,3) were infused for a period of 6 hours.
  • the administration of Compound A at 0.25 mg/kg/h prevented the development of hypotension (SEE FIG. 1) and greatly decreased mortality (20% mortality at 9h, n 10).
  • EXAMPLE 17 USE OF THE SOD MIMICS OF THE INVENTION AS ANTI-INFLAMMATORY AGENTS IN THE CARRAGEENAN-INDUCED PLEURISY INFLAMMATION MODEL
  • Male Sprague-Dawley rats 300-350 g; Charles River; Milan; Italy
  • Animal care was in compliance with Italian regulations on protection of animals used for experimental and other scientific purposes (DM. 116192) as well as with the EEC regulations (O.J. of E.C. L 358/1 12/18/1986). Rats were anaesthetised with isoflurane and submitted to a skin incision at the level ofthe left sixth intercostal space.
  • the underlying muscle was dissected and saline (0.2 ml) or saline containing 1% ⁇ -canageenan (0.2 ml), injected into the pleural cavity.
  • the skin incision was closed with a suture and the animals allowed to recover.
  • Compound 16, Compound 31, and Compound 25, (.5-20 mg/kg, as indicated), or an equivalent volume (0.3 ml) of vehicle (26 mM sodium bicarbonate buffer, pH 8.1-8.3), was injected intraperitoneally (i.p.) 15 min before canageenan.
  • vehicle 26 mM sodium bicarbonate buffer, pH 8.1-8.3
  • the chest was carefully opened and the pleural cavity rinsed with 2 ml of saline solution containing heparin (5 U/ml) and indomethacin (10 ⁇ g/ml).
  • the exudate and washing solution were removed by aspiration and the total volume measured. Any exudate, that was contaminated with blood was discarded. The amount of exudate was calculated by subtracting the volume injected (2 ml) from the total volume recovered.
  • the leukocytes in the exudate were suspended in phosphate-buffer saline (PBS) and counted with an optical microscope in a Burker's chamber after vital Trypan Blue staining. Measurement of lung-tissue myeloperoxidase activity and malondialdehyde.
  • PBS phosphate-buffer saline
  • MPO Myeloperoxidase activity
  • a hemoprotein located in azurophil granules of neutrophils has been used as a biochemical marker for neutropbil infiltration into tissues (Bradley et al., 1982).
  • MPO was measured photometrically by a method similar to that described previously (Laight et al., 1994).
  • lung tissues were obtained and weighed. Each piece of tissue was homogenized in a solution containing 0.5% hexa- decyl-trimethyl-ammonium bromide dissolved in 10 mM potassium phosphate buffer (pH 7) and centrifuged for 30 min. at 20,000 x g at 4°C.
  • MDA lipid peroxidation
  • Treatment of rats with Compound 16, Compound 31 and Compound 25, (.5-20 mg/kg given by intraperitoneal injection, i.p. 15 min prior to ca ⁇ ageenan, as indicated) attenuated all parameters of inflammation.
  • the results are set forth in the table below.
  • MAP mean arterial pressure
  • Compound A was infused at either 0.075 or 0.25 mg/Kg/h 3 hours after live E.
  • Compound A All animals were given antibiotics 30 min and 9 hours after live E. Coli was administered.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Addiction (AREA)
  • Toxicology (AREA)
  • Rheumatology (AREA)
  • Dermatology (AREA)
  • Epidemiology (AREA)
  • Pain & Pain Management (AREA)
  • Pulmonology (AREA)
  • Hematology (AREA)
  • Transplantation (AREA)
  • Ophthalmology & Optometry (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Vascular Medicine (AREA)
  • Biochemistry (AREA)
  • Urology & Nephrology (AREA)
  • Oncology (AREA)
  • Diabetes (AREA)
  • Psychiatry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
EP00966722A 1999-09-16 2000-09-14 Pyridino pentaazamacrocyle substitue possedant une activite de superoxyde dismutase Expired - Lifetime EP1212323B1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP04003746A EP1420022B1 (fr) 1999-09-16 2000-09-14 Utilisation de complexes de pyridinopentaazamacrocycle substitués à activité de superoxyde dismutase
EP04003751A EP1420019B1 (fr) 1999-09-16 2000-09-14 Ligans pyridino pentaazamacrocycliques substitués

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US09/398,120 US6214817B1 (en) 1997-06-20 1999-09-16 Substituted pyridino pentaazamacrocyle complexes having superoxide dismutase activity
US398120 1999-09-16
PCT/US2000/025154 WO2001019823A2 (fr) 1999-09-16 2000-09-14 Pyridino pentaazamacrocyle substitue possedant une activite de superoxyde dismutase

Related Child Applications (2)

Application Number Title Priority Date Filing Date
EP04003751A Division EP1420019B1 (fr) 1999-09-16 2000-09-14 Ligans pyridino pentaazamacrocycliques substitués
EP04003746A Division EP1420022B1 (fr) 1999-09-16 2000-09-14 Utilisation de complexes de pyridinopentaazamacrocycle substitués à activité de superoxyde dismutase

Publications (2)

Publication Number Publication Date
EP1212323A2 true EP1212323A2 (fr) 2002-06-12
EP1212323B1 EP1212323B1 (fr) 2004-06-09

Family

ID=23574065

Family Applications (3)

Application Number Title Priority Date Filing Date
EP04003746A Expired - Lifetime EP1420022B1 (fr) 1999-09-16 2000-09-14 Utilisation de complexes de pyridinopentaazamacrocycle substitués à activité de superoxyde dismutase
EP00966722A Expired - Lifetime EP1212323B1 (fr) 1999-09-16 2000-09-14 Pyridino pentaazamacrocyle substitue possedant une activite de superoxyde dismutase
EP04003751A Expired - Lifetime EP1420019B1 (fr) 1999-09-16 2000-09-14 Ligans pyridino pentaazamacrocycliques substitués

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP04003746A Expired - Lifetime EP1420022B1 (fr) 1999-09-16 2000-09-14 Utilisation de complexes de pyridinopentaazamacrocycle substitués à activité de superoxyde dismutase

Family Applications After (1)

Application Number Title Priority Date Filing Date
EP04003751A Expired - Lifetime EP1420019B1 (fr) 1999-09-16 2000-09-14 Ligans pyridino pentaazamacrocycliques substitués

Country Status (12)

Country Link
US (1) US6214817B1 (fr)
EP (3) EP1420022B1 (fr)
JP (2) JP2003509423A (fr)
AT (3) ATE368039T1 (fr)
AU (1) AU784078B2 (fr)
CA (1) CA2382105C (fr)
DE (3) DE60011446T2 (fr)
DK (1) DK1212323T3 (fr)
ES (3) ES2293106T3 (fr)
HK (1) HK1046689B (fr)
PT (1) PT1212323E (fr)
WO (1) WO2001019823A2 (fr)

Families Citing this family (68)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6127356A (en) 1993-10-15 2000-10-03 Duke University Oxidant scavengers
EP0846003A1 (fr) * 1995-08-17 1998-06-10 Monsanto Company Methodes d'analyse d'images diagnostiques utilisant des conjugues biologiques de complexes metalliques de ligands macrocycliques renfermant de l'azote
US20050171198A1 (en) * 1997-06-20 2005-08-04 Metaphore Pharmaceuticals, Inc. SODm therapy for treatment, prevention, inhibition and reversal of inflammatory disease
EP1045851B1 (fr) 1997-11-03 2003-04-23 Duke University Porphyrines substituees
ES2385486T3 (es) 1999-01-25 2012-07-25 National Jewish Health Porfirinas sustituidas y su uso terapéutico
US20030069281A1 (en) * 2000-06-14 2003-04-10 Irwin Fridovich Tetrapyrroles
US20020072512A1 (en) * 2000-12-08 2002-06-13 Metaphore Pharmaceuticals, Inc Method of preventing and treating HIV-mediated central nervous system damage
DE60235955D1 (de) * 2001-01-05 2010-05-27 Metaphore Pharmaceuticals Inc Zusammensetzungen und verfahren zur erhöhung der zytokin-aktivität und zur behandlung der hypotension assoziiert mit der verabreichung von zytokinen
DE60233317D1 (de) 2001-01-19 2009-09-24 Nat Jewish Med & Res Center Medikament zum schutz in der radiotherapie
WO2002058686A2 (fr) * 2001-01-26 2002-08-01 Metaphore Pharmaceuticals, Inc. Methode de traitement de troubles neurodegeneratifs
US20040137638A1 (en) * 2002-03-04 2004-07-15 Slomczynska Urszula J. Chromatography of metal complexes
CA2439971A1 (fr) * 2001-03-02 2002-09-12 Metaphore Pharmaceuticals, Inc. Chromatographie de complexes metalliques
EP1439842A4 (fr) * 2001-06-01 2009-09-02 Nat Jewish Med & Res Center Capteurs oxydants destines au traitement du diabete ou a etre utilises dans une transplantation ou a induire une tolerance immunitaire
AU2002315001A1 (en) * 2001-06-08 2002-12-23 Metaphore Pharmaceuticals, Inc. Libraries of conformationally constrained peptides, chiral azacrowns, and peptidomimetics and methods of making the same
MXPA03011795A (es) * 2001-06-26 2005-10-18 Metaphore Pharmaceuticals Inc Terapia de combinacion de una superoxido dismutasa mimetica y un corticosteroide para la prevencion y/o el tratamiento de una enfermedad inflamatoria.
US20040132706A1 (en) * 2001-10-05 2004-07-08 Daniela Salvemini Composition comprising a catalyst for the dismutation of superoxide and use of the composition for preventing and treating hypotension
KR100951606B1 (ko) * 2001-12-14 2010-04-09 알콘 인코포레이티드 안과 질환 및 질병의 치료를 위한 과산화물 디스뮤타제모방체
EP1513537A4 (fr) * 2002-06-07 2006-09-06 Univ Duke Porphyrines substituees
US6835387B2 (en) * 2002-06-11 2004-12-28 Scimed Life Systems, Inc. Sustained release of superoxide dismutase mimics from implantable or insertable medical devices
US20030232784A1 (en) * 2002-06-14 2003-12-18 Benedict James J. Hyaluronic acid-associated superoxide dismutase mimics and their use in the treatment of joint pain
US6984636B2 (en) * 2002-08-12 2006-01-10 Medical Research Council Mitochondrially targeted antioxidants
WO2004050084A2 (fr) * 2002-11-29 2004-06-17 Yissum Research Development Company Of The Hebrew University Of Jerusalem Inhibiteurs d'enzyme de conversion de l'angiotensine possedant une activite antioxydante et de donneur d'oxyde nitrique (no)
MXPA05005937A (es) * 2002-12-06 2005-08-18 Alcon Inc Imitaciones de superoxido de dismutasa para el tratamiento de trastornos y enfermedades oculares.
EP1581212A4 (fr) * 2002-12-06 2008-11-05 Alcon Inc Analogues de dismutase de superoxyde destines au traitement de troubles et maladies oculaires
US7299082B2 (en) 2003-10-31 2007-11-20 Abbott Diabetes Care, Inc. Method of calibrating an analyte-measurement device, and associated methods, devices and systems
JP2007513948A (ja) * 2003-12-11 2007-05-31 アルコン，インコーポレイテッド 視神経損傷および網膜損傷の処置のためのスーパーオキシドジスムターゼ模倣物
FR2863892B1 (fr) * 2003-12-18 2007-05-11 Univ Paris Descartes Utilisation de mimetiques de la superoxyde dismutase et de la glutathion reductase comme anti-cancereux
US7699964B2 (en) 2004-02-09 2010-04-20 Abbott Diabetes Care Inc. Membrane suitable for use in an analyte sensor, analyte sensor, and associated method
US8165651B2 (en) * 2004-02-09 2012-04-24 Abbott Diabetes Care Inc. Analyte sensor, and associated system and method employing a catalytic agent
US8217166B2 (en) * 2005-05-05 2012-07-10 Galera Therapeutics, Llc Polyethylene glycolated superoxide dismutase mimetics
US20120301403A1 (en) 2007-06-21 2012-11-29 Galera Therapeutics, Llc Polyethylene glycolated superoxide dismutase mimetics
KR100724586B1 (ko) 2005-12-09 2007-06-04 세종대학교산학협력단 수퍼옥사이드 디스뮤타아제 활성을 갖는 고리화합물
US7885698B2 (en) 2006-02-28 2011-02-08 Abbott Diabetes Care Inc. Method and system for providing continuous calibration of implantable analyte sensors
EP3527079A1 (fr) 2006-10-12 2019-08-21 Galera Labs, LLC Procédés de traitement de la mucosite orale
WO2008114017A2 (fr) * 2007-03-19 2008-09-25 Imperial Innovations Limited Compositions et procédés associés à l'infection par le virus de l'influenza
WO2009143454A2 (fr) 2008-05-22 2009-11-26 Kereos, Inc. Combinaison thérapeutique anticancéreuse
KR101646066B1 (ko) 2008-05-23 2016-08-05 내셔날 쥬이쉬 헬스 알킬화 종으로의 노출과 관련된 상해를 치료하는 방법
PL2321364T3 (pl) 2008-08-22 2016-01-29 Novomer Inc Sposoby syntezy polimerów
AU2009333303A1 (en) * 2008-12-17 2011-08-11 Altheus Therapeutics, Inc. Enema formulations
CA2849456C (fr) * 2011-09-26 2021-12-07 Galera Therapeutics, Llc Methodes de traitement de maladies
HUE035069T2 (en) 2011-12-28 2018-05-02 Global Blood Therapeutics Inc Substituted benzaldehyde compounds and their use to increase tissue oxygenation
WO2013102145A1 (fr) 2011-12-28 2013-07-04 Global Blood Therapeutics, Inc. Composés d'aldéhydes hétéroaryles substitués et leurs procédés d'utilisation dans l'accroissement de l'oxygénation tissulaire
US8952171B2 (en) 2013-03-15 2015-02-10 Global Blood Therapeutics, Inc. Compounds and uses thereof for the modulation of hemoglobin
US9604999B2 (en) 2013-03-15 2017-03-28 Global Blood Therapeutics, Inc. Compounds and uses thereof for the modulation of hemoglobin
US9802900B2 (en) 2013-03-15 2017-10-31 Global Blood Therapeutics, Inc. Bicyclic heteroaryl compounds and uses thereof for the modulation of hemoglobin
CA2903022C (fr) 2013-03-15 2021-11-09 Global Blood Therapeutics, Inc. Composes et leurs utilisations pour la modulation de l'hemoglobine
US20140274961A1 (en) 2013-03-15 2014-09-18 Global Blood Therapeutics, Inc. Compounds and uses thereof for the modulation of hemoglobin
US9458139B2 (en) 2013-03-15 2016-10-04 Global Blood Therapeutics, Inc. Compounds and uses thereof for the modulation of hemoglobin
US9422279B2 (en) 2013-03-15 2016-08-23 Global Blood Therapeutics, Inc. Compounds and uses thereof for the modulation of hemoglobin
PE20161035A1 (es) 2013-03-15 2016-11-13 Global Blood Therapeutics Inc Compuestos y usos de estos para la modulacion de la hemoglobina
KR20150132146A (ko) 2013-03-15 2015-11-25 글로벌 블러드 테라퓨틱스, 인크. 헤모글로빈 조정을 위한 화합물 및 이의 용도
US10100043B2 (en) 2013-03-15 2018-10-16 Global Blood Therapeutics, Inc. Substituted aldehyde compounds and methods for their use in increasing tissue oxygenation
US10266551B2 (en) 2013-03-15 2019-04-23 Global Blood Therapeutics, Inc. Compounds and uses thereof for the modulation of hemoglobin
EA201992707A1 (ru) 2013-11-18 2020-06-30 Глобал Блад Терапьютикс, Инк. Соединения и их применения для модуляции гемоглобина
WO2015102745A1 (fr) 2013-12-31 2015-07-09 Abbott Diabetes Care Inc. Capteur d'analytes auto-alimenté et dispositifs utilisant ce dernier
JP6809681B2 (ja) 2014-02-07 2021-01-06 グローバル ブラッド セラピューティクス インコーポレイテッド ２−ヒドロキシ−６−（（２−（１−イソプロピル−１ｈ−ピラゾール−５−イル）ピリジン−３−イル）メトキシ）ベンズアルデヒドの遊離塩基の結晶多形
MA41841A (fr) 2015-03-30 2018-02-06 Global Blood Therapeutics Inc Composés aldéhyde pour le traitement de la fibrose pulmonaire, de l'hypoxie, et de maladies auto-immunes et des tissus conjonctifs
AU2016306568B2 (en) * 2015-08-11 2021-01-07 Galera Labs, Llc Pentaaza macrocyclic ring complexes possessing oral bioavailability
TW201731509A (zh) 2015-12-04 2017-09-16 全球血液治療公司 針對２-羥基-６-（（２-（１-異丙基-１ｈ-吡唑-５-基）吡啶-３-基）甲氧基）-苯甲醛之劑量方案
TWI663160B (zh) 2016-05-12 2019-06-21 全球血液治療公司 用於合成2-羥基-6-((2-(1-異丙基-1h-吡唑-5-基)-吡啶-3-基)甲氧基)苯甲醛之方法
JP7166634B2 (ja) * 2016-09-01 2022-11-08 ガレラ・ラブス・リミテッド・ライアビリティ・カンパニー ペンタアザ大環状環複合体およびアスコルビン酸化合物による組み合わせ癌療法
TW202332423A (zh) 2016-10-12 2023-08-16 美商全球血液治療公司 包含2-羥基-6-((2-(1-異丙基-1h-吡唑-5-基)吡啶-3-基)甲氧基)-苯甲醛之片劑
WO2018152353A2 (fr) 2017-02-15 2018-08-23 Galera Labs, Llc Complexes de noyaux pentaaza-macrocycliques destinés à une administration intestinale locale
US11246950B2 (en) 2017-04-13 2022-02-15 Galera Labs, Llc Combination cancer immunotherapy with pentaaza macrocyclic ring complex
SG11201909495PA (en) * 2017-04-13 2019-11-28 Galera Labs Llc Combination cancer immunotherapy with pentaaza macrocyclic ring complex
EA202091832A1 (ru) 2018-01-31 2021-01-11 ГАЛЕРА ЛЭБЗ, ЭлЭлСи Комбинированная терапия злокачественных новообразований макроциклическим кольцевым комплексом пентаазы и противоопухолевым агентом на основе платины
EP3860975B1 (fr) 2018-10-01 2023-10-18 Global Blood Therapeutics, Inc. Modulateurs de l'hémoglobine pour le traitement de la drépanocytose
CN115260178B (zh) * 2022-08-09 2024-09-20 上海师范大学 一种吡啶骨架ncn噁唑啉类配体的合成方法和应用

Family Cites Families (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4001212A (en) 1973-10-03 1977-01-04 E. I. Du Pont De Nemours And Company Macrocyclic polyamines
US3930867A (en) 1974-01-07 1976-01-06 E. I. Du Pont De Nemours And Company Macrocyclic polyamines as sensitizers for silver halide emulsions
NO149961C (no) 1981-06-01 1984-07-25 Borregaard Ind Fremgangsmaate for fremstilling av n-(2-hydroksyetyl)-derivater av makrocykliske polyaminer, inneholdende flere nitrogenatomer i 1,4-forhold i ringen
JPS61226751A (ja) 1985-03-30 1986-10-08 Konishiroku Photo Ind Co Ltd ハロゲン化銀写真感光材料用処理液及びその処理方法
US4968616A (en) 1986-05-16 1990-11-06 Masayasu Inoue Superoxide dismutase derivatives, method of producing same and medicinal use of same
US5096724A (en) 1988-06-03 1992-03-17 Aquanautics Corporation Methods, compositions, and systems for ligand extraction
US5252720A (en) 1989-03-06 1993-10-12 Board Of Regents, The University Of Texas System Metal complexes of water soluble texaphyrins
GB9001245D0 (en) 1990-01-19 1990-03-21 Salutar Inc Compounds
EP0511275A1 (fr) 1990-01-19 1992-11-04 Nycomed Imaging As Composes de chelation
GB9024208D0 (en) 1990-11-07 1990-12-19 Salutar Inc Compounds
KR0145953B1 (ko) * 1991-07-19 1998-08-17 제임스 클리프튼 보올딩 슈퍼옥사이드의 불균화촉매로서 효과적인 질소를 포함하는 마크로사이클 릭리간드의 망간착물
CA2072934C (fr) 1991-07-19 2007-08-28 Karl William Aston Complexes de manganese et de ligands macrocycliques contenant de l'azote, efficaces comme catalyseurs de la dismutation de superoxydes
WO1993011800A1 (fr) 1991-12-10 1993-06-24 The Dow Chemical Company Complexes et conjugues a base d'acides bicyclopolyazamacrocyclocarboxylique, leur preparation et utilisation et tant qu'agents de contraste
US5403834A (en) 1992-12-07 1995-04-04 Eukarion, Inc. Synthetic catalytic free radical scavengers useful as antioxidants for prevention and therapy of disease
US5696109A (en) 1992-12-07 1997-12-09 Eukarion, Inc. Synthetic catalytic free radical scavengers useful as antioxidants for prevention and therapy of disease
US5834509A (en) 1992-12-07 1998-11-10 Eukarion, Inc. Synthetic catalytic free radical scavengers useful as antioxidants for prevention and therapy of disease
US6204259B1 (en) 1993-01-14 2001-03-20 Monsanto Company Manganese complexes of nitrogen-containing macrocyclic ligands effective as catalysts for dismutating superoxide
WO1995028968A1 (fr) 1994-04-22 1995-11-02 Monsanto Company Analyse d'image de diagnostic utilisant des complexes metalliques
WO1996039409A1 (fr) 1995-06-05 1996-12-12 Nitromed, Inc. Oxydants et reducteurs nitrosyles et nitres de superoxydes
US6525041B1 (en) 1995-06-06 2003-02-25 Pharmacia Corporation Manganese or iron complexes of nitrogen-containing macrocyclic ligands effective as catalysts for dismutating superoxide
JPH11507621A (ja) 1995-06-07 1999-07-06 モンサント カンパニー 置換ポリアザ大環状化合物の製造法
SE509595C2 (sv) * 1996-01-25 1999-02-15 Sandvik Ab Skär för spånavskiljande metallbearbetning
BR9708179A (pt) * 1996-03-13 1999-07-27 Monsanto Co Bioconjugados de complexos de manganês ou ferro de ligantes macrociclicos contendo nitrogênio eficazes como catalizadores para desmutar superoxidos
US6180620B1 (en) 1997-06-20 2001-01-30 G.D. Searle & Co. Analgesic methods using synthetic catalysts for the dismutation of superoxide radicals

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0119823A2 *

Also Published As

Publication number Publication date
CA2382105A1 (fr) 2001-03-22
CA2382105C (fr) 2012-01-10
WO2001019823A3 (fr) 2001-09-07
DE60011446D1 (de) 2004-07-15
WO2001019823A2 (fr) 2001-03-22
EP1420022B1 (fr) 2007-07-04
ES2222925T3 (es) 2005-02-16
ATE366254T1 (de) 2007-07-15
EP1420019B1 (fr) 2007-07-25
AU7702400A (en) 2001-04-17
ES2293106T3 (es) 2008-03-16
DE60035724D1 (de) 2007-09-06
WO2001019823A9 (fr) 2002-09-26
EP1420022A1 (fr) 2004-05-19
AU784078B2 (en) 2006-02-02
PT1212323E (pt) 2004-10-29
DE60011446T2 (de) 2005-07-07
DK1212323T3 (da) 2004-10-25
EP1420019A1 (fr) 2004-05-19
DE60035440T2 (de) 2008-03-13
DE60035724T2 (de) 2008-04-30
ES2291759T3 (es) 2008-03-01
JP2003509423A (ja) 2003-03-11
DE60035440D1 (de) 2007-08-16
HK1046689B (zh) 2005-03-11
HK1046689A1 (en) 2003-01-24
ATE268774T1 (de) 2004-06-15
JP2012097097A (ja) 2012-05-24
ATE368039T1 (de) 2007-08-15
EP1212323B1 (fr) 2004-06-09
US6214817B1 (en) 2001-04-10

Similar Documents

Publication Publication Date Title
EP1212323B1 (fr) Pyridino pentaazamacrocyle substitue possedant une activite de superoxyde dismutase
Adamovich New atranes and similar ionic complexes. Synthesis, structure, properties
US5637578A (en) Manganese complexes of nitrogen-containing macrocyclic ligands effective as catalysts for dismutating superoxide
AU780118B2 (en) Peroxynitrite decomposition catalysts and methods of use thereof
EP0598753B1 (fr) Complexes manganeseux de ligands macrocycliques azotes efficaces comme catalyseurs pour dismuter le superoxyde
US6525041B1 (en) Manganese or iron complexes of nitrogen-containing macrocyclic ligands effective as catalysts for dismutating superoxide
CA2151262A1 (fr) Complexes de manganese de ligands macrocycliques azotes, efficaces comme catalyseurs pour la dismutation de superoxydes
EA010834B1 (ru) Пиридилзамещённые порфириновые соединения и способы их применения
US8334377B2 (en) Porphyrin catalysts and methods of use thereof
Kostenkova et al. Vanadium (V) Pyridine‐Containing Schiff Base Catecholate Complexes are Lipophilic, Redox‐Active and Selectively Cytotoxic in Glioblastoma (T98G) Cells
US6589966B1 (en) Cytotoxic metal chelators and methods for making and using same
EP2545064B1 (fr) Composés de rhénium libérant du monoxyde de carbone à usage médical
US20240124501A1 (en) Vanadium compounds and methods of making and using thereof
Abdelbaste et al. Synthesis and in vitro biological examination of dioxovanadium (v) mixed-ligand complexes containing 4-amino-5-mercapto-3-phenyl-1, 2, 4-triazole as primary Ligand and o-aniline derivatives as co-ligands
Lakin Complexation of early-and post-transition metal halides with aza-, oxa-, and thiamacrocyclic ligands

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20020304

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

AX Request for extension of the european patent

Free format text: AL;LT;LV;MK;RO;SI

17Q First examination report despatched

Effective date: 20020911

RIN1 Information on inventor provided before grant (corrected)

Inventor name: LENNON, PATRICK

Inventor name: SIKORSKI, JAMES, A.

Inventor name: SALVEMINI, DANIELA

Inventor name: HENKE, SUSAN, L.

Inventor name: FOBIAN, YVETTE

Inventor name: NEUMANN, WILLIAM, L.

Inventor name: KUSTURIN, CARRIE, L.

Inventor name: GRAPPERHAUS, MARGARET, L.

Inventor name: RILEY, DENNIS

Inventor name: ASTON, KARL, W.

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

REG Reference to a national code

Ref country code: GB

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: CH

Ref legal event code: EP

REF Corresponds to:

Ref document number: 60011446

Country of ref document: DE

Date of ref document: 20040715

Kind code of ref document: P

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: MC

Payment date: 20040826

Year of fee payment: 5

REG Reference to a national code

Ref country code: CH

Ref legal event code: NV

Representative=s name: BOVARD AG PATENTANWAELTE

REG Reference to a national code

Ref country code: SE

Ref legal event code: TRGR

REG Reference to a national code

Ref country code: GR

Ref legal event code: EP

Ref document number: 20040402959

Country of ref document: GR

REG Reference to a national code

Ref country code: DK

Ref legal event code: T3

REG Reference to a national code

Ref country code: PT

Ref legal event code: SC4A

Free format text: AVAILABILITY OF NATIONAL TRANSLATION

Effective date: 20040907

LTIE Lt: invalidation of european patent or patent extension

Effective date: 20040609

REG Reference to a national code

Ref country code: ES

Ref legal event code: FG2A

Ref document number: 2222925

Country of ref document: ES

Kind code of ref document: T3

ET Fr: translation filed
REG Reference to a national code

Ref country code: HK

Ref legal event code: GR

Ref document number: 1046689

Country of ref document: HK

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed

Effective date: 20050310

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GR

Payment date: 20050826

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CY

Payment date: 20050831

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 20050904

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: SE

Payment date: 20050906

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: AT

Payment date: 20050913

Year of fee payment: 6

Ref country code: IE

Payment date: 20050913

Year of fee payment: 6

Ref country code: PT

Payment date: 20050913

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 20050914

Year of fee payment: 6

Ref country code: FI

Payment date: 20050914

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DK

Payment date: 20050915

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: LU

Payment date: 20050928

Year of fee payment: 6

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MC

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050930

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: BE

Payment date: 20051125

Year of fee payment: 6

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060914

Ref country code: FI

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060914

Ref country code: AT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060914

Ref country code: CY

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060914

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060915

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: CH

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060930

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060930

Ref country code: LI

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060930

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DK

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20061002

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: PT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20070314

REG Reference to a national code

Ref country code: PT

Ref legal event code: MM4A

Free format text: LAPSE DUE TO NON-PAYMENT OF FEES

Effective date: 20070314

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NL

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20070401

REG Reference to a national code

Ref country code: DK

Ref legal event code: EBP

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

EUG Se: european patent has lapsed
NLV4 Nl: lapsed or anulled due to non-payment of the annual fee

Effective date: 20070401

REG Reference to a national code

Ref country code: IE

Ref legal event code: MM4A

BERE Be: lapsed

Owner name: *MONSANTO CY

Effective date: 20060930

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20060914

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20070404

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20140929

Year of fee payment: 15

Ref country code: FR

Payment date: 20140917

Year of fee payment: 15

Ref country code: ES

Payment date: 20140926

Year of fee payment: 15

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: IT

Payment date: 20140924

Year of fee payment: 15

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20140929

Year of fee payment: 15

REG Reference to a national code

Ref country code: DE

Ref legal event code: R119

Ref document number: 60011446

Country of ref document: DE

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20150914

GBPC Gb: european patent ceased through non-payment of renewal fee

Effective date: 20150914

REG Reference to a national code

Ref country code: FR

Ref legal event code: ST

Effective date: 20160531

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20160401

Ref country code: GB

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20150914

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: FR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20150930

REG Reference to a national code

Ref country code: ES

Ref legal event code: FD2A

Effective date: 20161028

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: ES

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20150915